Novel garden bean sb4355

ABSTRACT

A novel garden bean cultivar, designated SB4355, is disclosed. The invention relates to the seeds of garden bean cultivar SB4355, to the plants of garden bean line SB4355 and to methods for producing a garden bean plant by crossing the cultivar SB4355 with itself or another garden bean line. The invention further relates to methods for producing a garden bean plant containing in its genetic material one or more transgenes and to the transgenic plants produced by that method and to methods for producing other garden bean lines derived from the cultivar SB4355.

FIELD OF THE INVENTION

The present invention relates to a new and distinctive Garden Bean(Phaseolus vulgaris) cultivar, designated SB4355.

BACKGROUND OF THE INVENTION

There are numerous steps in the development of any novel, desirableplant germplasm. Plant breeding begins with the analysis and definitionof problems and weaknesses of the current germplasm, the establishmentof program goals, and the definition of specific breeding objectives.The next step is selection of germplasm that possess the traits to meetthe program goals. The goal is to combine in a single cultivar or hybridan improved combination of desirable traits from the parental germplasm.These important traits may include fresh pod yield, higher seed yield,resistance to diseases and insects, better stems and roots, tolerance todrought and heat, and better agronomic quality. With mechanicalharvesting of many crops, uniformity of plant characteristics such asgermination and stand establishment, growth rate, maturity and plantheight is important.

Choice of breeding or selection methods depends on the mode of plantreproduction, the heritability of the trait(s) being improved, and thetype of cultivar used commercially (e.g., F1 hybrid cultivar, purelinecultivar, etc.). For highly heritable traits, a choice of superiorindividual plants evaluated at a single location will be effective,whereas for traits with low heritability, selection should be based onmean values obtained from replicated evaluations of families of relatedplants. Popular selection methods commonly include pedigree selection,modified pedigree selection, mass selection, and recurrent selection.

The complexity of inheritance influences choice of the breeding method.Backcross breeding is used to transfer one or a few favorable genes fora highly heritable trait into a desirable cultivar. This approach hasbeen used extensively for breeding disease-resistant cultivars. Variousrecurrent selection techniques are used to improve quantitativelyinherited traits controlled by numerous genes. The use of recurrentselection in self-pollinating crops depends on the ease of pollination,the frequency of successful hybrids from each pollination, and thenumber of hybrid offspring from each successful cross.

Each breeding program should include a periodic, objective evaluation ofthe efficiency of the breeding procedure. Evaluation criteria varydepending on the goal and objectives, but should include gain fromselection per year based on comparisons to an appropriate standard,overall value of the advanced breeding lines, and number of successfulcultivars produced per unit of input (e.g., per year, per dollarexpended etc.).

Promising advanced breeding lines are thoroughly tested and compared toappropriate standards in environments representative of the commercialtarget area(s) for at least three years. The best lines are candidatesfor new commercial cultivars; those still deficient in a few traits areused as parents to produce new populations for further selection.

These processes, which lead to the final step of marketing anddistribution, usually take from eight to twelve years from the time thefirst cross is made. Therefore, development of new cultivars is atime-consuming process that requires precise forward planning, efficientuse of resources, and a minimum of changes in direction.

A most difficult task is the identification of individuals that aregenetically superior, because for most traits the true genotypic valueis masked by other confounding plant traits or environmental factors.One method of identifying a superior plant is to observe its performancerelative to other experimental plants and to a widely grown standardcultivar. If a single observation is inconclusive, replicatedobservations provide a better estimate of its genetic worth.

The goal of plant breeding is to develop new, unique and superior gardenbean cultivars. The breeder initially selects and crosses two or moreparental lines, followed by repeated selfing and selection, producingmany new genetic combinations. The breeder can theoretically generatebillions of different genetic combinations via crossing, selfing andmutations. The breeder has no direct control at the cellular level.Therefore, two breeders will never develop the same line. Each year, theplant breeder selects the germplasm to advance to the next generation.This germplasm is grown under unique and different geographical,climatic and soil conditions, and further selections are then made,during and at the end of the growing season. The cultivars that aredeveloped are unpredictable. This unpredictability is because thebreeder's selection occurs in unique environments, with no control atthe DNA level (using conventional breeding procedures), and withmillions of different possible genetic combinations being generated. Abreeder of ordinary skill in the art cannot predict the final resultinglines he develops, except possibly in a very gross and general fashion.The same breeder cannot produce the same line twice by using the exactsame original parents and the same selection techniques. Thisunpredictability results in the expenditure of large research monies todevelop superior garden bean cultivars.

The development of commercial garden bean cultivars requires thedevelopment of garden bean varieties, the crossing of these varieties,and the evaluation of the crosses. Pedigree breeding and recurrentselection breeding methods are used to develop cultivars from breedingpopulations. Breeding programs combine desirable traits from two or morevarieties or various broad-based sources into breeding pools from whichcultivars are developed by selfing and selection of desired phenotypes.The new cultivars are crossed with other varieties and the progeny fromthese crosses are evaluated to determine which have commercial potentialas a new cultivar.

Pedigree breeding is used commonly for the improvement ofself-pollinating crops or inbred lines of cross-pollinating crops. Twoparents that possess favorable, complementary traits are crossed toproduce an F1. An F2 population is produced by selfing one or severalF1's or by intercrossing two F1's (sib mating). Selection of the bestindividuals is usually begun in the F2 population; then, beginning inthe F3, the best individuals in the best families are selected.Replicated testing of families, or hybrid combinations involvingindividuals of these families, often follows in the F4 generation toimprove the effectiveness of selection for traits with low heritability.At an advanced stage of inbreeding (i.e., F6 and F7), the best lines ormixtures of phenotypically similar lines are tested for potentialrelease as new cultivars. Mass and recurrent selections can be used toimprove populations of either self- or cross-pollinating crops. Agenetically variable population of heterozygous individuals is eitheridentified or created by intercrossing several different parents. Thebest plants are selected based on individual superiority, outstandingprogeny, or excellent combining ability. The selected plants areintercrossed to produce a new population in which further cycles ofselection are continued.

Backcross breeding has been used to transfer genes for a simplyinherited, highly heritable trait into a desirable homozygous cultivaror line that is the recurrent parent. The source of the trait to betransferred is called the donor parent. The resulting plant is expectedto have the attributes of the recurrent parent (e.g., cultivar) and thedesirable trait transferred from the donor parent. After the initialcross, individuals possessing the phenotype of the donor parent areselected and repeatedly crossed (backcrossed) to the recurrent parent.The resulting plant is expected to have the attributes of the recurrentparent (e.g., cultivar) and the desirable trait transferred from thedonor parent.

The single-seed descent procedure in the strict sense refers to plantinga segregating population, harvesting a sample of one seed per plant, andusing the one-seed sample to plant the next generation. When thepopulation has been advanced from the F2 to the desired level ofinbreeding, the plants from which lines are derived will each trace todifferent F2 individuals. The number of plants in a population declineseach generation due to failure of some seeds to germinate or some plantsto produce at least one seed. As a result, not all of the F2 plantsoriginally sampled in the population will be represented by a progenywhen generation advance is completed.

Descriptions of other breeding methods that are commonly used fordifferent traits and crops can be found in one of several referencebooks (e.g., “Principles of Plant Breeding” John Wiley and Son, pp.115-161, 1960; Allard, 1960; Simmonds, 1979; Sneep et al., 1979; Fehr,1987). Proper testing should detect any major faults and establish thelevel of superiority or improvement over current cultivars. In additionto showing superior performance, there must be a demand for a newcultivar that is compatible with industry standards or which creates anew market. The introduction of a new cultivar may incur additionalcosts to the seed producer, the grower, processor and consumer; forspecial advertising and marketing, altered seed and commercialproduction practices, and new product utilization. The testing precedingrelease of a new cultivar should take into consideration research anddevelopment costs as well as technical superiority of the finalcultivar. For seed-propagated cultivars, it must be feasible to produceseed easily and economically.

Garden bean, Phaseolus vulgaris L., is an important and valuablevegetable crop. Thus, a continuing goal of plant breeders is to developstable, high yielding garden bean cultivars that are agronomicallysound. The reasons for this goal are obviously to maximize the amount ofyield produced on the land. To accomplish this goal, the garden beanbreeder must select and develop garden bean plants that have the traitsthat result in superior cultivars.

SUMMARY OF THE INVENTION

According to the invention, there is provided a novel garden beancultivar, designated SB4355. This invention thus relates to the seeds ofgarden bean cultivar SB4355, to the plants of garden bean cultivarSB4355 and parts thereof, for example pollen, ovule or pod, and tomethods for producing a garden bean plant produced by crossing thegarden bean SB4355 with itself or another garden bean line, and tomethods for producing a garden bean plant containing in its geneticmaterial one or more transgenes and to the transgenic garden bean plantsproduced by that method. This invention also relates to methods forproducing other garden bean cultivars derived from garden bean cultivarSB4355 and to the garden bean cultivar derived by the use of thosemethods. This invention further relates to hybrid garden bean seeds andplants produced by crossing the line SB4355 with another garden beanline.

The invention is also directed to a method of producing a pod comprisinggrowing a plant according to the instant invention to produce a pod, andharvesting said pod. In one embodiment, the method further comprisesprocessing said pod to obtain a bean product. In one embodiment, a beanproduct according the instant invention is a fresh produce, a cannedproduct or a frozen product.

The invention is also directed to a method of producing a berrycomprising obtaining a pod of a plant according to the instant inventionand processing the pod to obtain a berry. In one embodiment, a berryaccording the instant invention is a fresh product, a canned product ora frozen product.

In another aspect, the present invention provides regenerable cells foruse in tissue culture of garden bean cultivar SB4355. The tissue culturewill preferably be capable of regenerating plants having thephysiological and morphological characteristics of the foregoing gardenbean plant, and of regenerating plants having substantially the samegenotype as the foregoing garden bean plant. Preferably, the regenerablecells in such tissue cultures will be embryos, protoplasts, seeds,callus, pollen, leaves, anthers, roots, and meristematic cells. Stillfurther, the present invention provides garden bean plants regeneratedfrom the tissue cultures of the invention.

Another objective of the invention is to provide methods for producingother garden bean plants derived from garden bean cultivar SB4355.Garden bean cultivars derived by the use of those methods are also partof the invention.

The invention also relates to methods for producing a garden bean plantcontaining in its genetic material one or more transgenes and to thetransgenic garden bean plant produced by that method.

In another aspect, the present invention provides for single geneconverted plants of SB4355. The single transferred gene may preferablybe a dominant or recessive allele. Preferably, the single transferredgene will confer such trait as male sterility, herbicide resistance,insect resistance, resistance for bacterial, fungal, or viral disease,male fertility, enhanced nutritional quality and industrial usage. Thesingle gene may be a naturally occurring garden bean gene or a transgeneintroduced through genetic engineering techniques.

The invention further provides methods for developing a garden beanplant in a garden bean plant breeding program using plant breedingtechnique including recurrent selection, backcrossing, pedigreebreeding, restriction fragment length polymorphism enhanced selection,genetic marker enhanced selection and transformation. Seeds, garden beanplant, and parts thereof produced by such breeding methods are also partof the invention.

Definitions

In the description and tables, which follow, a number of terms are used.In order to provide a clear and consistent understanding of thespecification and claims, including the scope to be given such terms,the following definitions are provided:

-   “Allele”—The allele is any of one or more alternative form of a    gene, all of which alleles relates to one trait or characteristic.    In a diploid cell or organism, the two alleles of a given gene    occupy corresponding loci on a pair of homologous chromosomes.-   “Backcrossing”—Backcrossing is a process in which a breeder    repeatedly crosses hybrid progeny back to one of the parents, for    example, a first generation hybrid F1 with one of the parental    genotype of the F1 hybrid.-   “Essentially all the physiological and morphological    characteristics”—A plant having essentially all the physiological    and morphological characteristics means a plant having the    physiological and morphological characteristics, except for the    characteristics derived from the converted gene.-   “Regeneration”—Regeneration refers to the development of a plant    from tissue culture.-   “Single gene converted”—Single gene converted or conversion plant    refers to plants which are developed by a plant breeding technique    called backcrossing wherein essentially all of the desired    morphological and physiological characteristics of a line are    recovered in addition to the single gene transferred into the line    via the backcrossing technique or via genetic engineering.-   “Maturity Date”—Plants are considered mature when the pods have    reached their maximum desirable seed size and sieve size for the    specific use intended.-   “Determinate Plant”—a determinate plant will grow to a fixed number    of nodes while an indeterminate plant will continue to grow during    the season. They have a high pod to vine weight ratio.-   “Sieve Size”—(sv)—Sieve size measures the diameter of the fresh pod    and is used in grading beans. Sieve size 1 means pods that fall    through a sieve grader which culls out pod diameters of 4.76 mm    through 5.76 mm. Sieve size 2 means pods that fall through a sieve    grader which culls out pod diameters of 5.76 mm through 7.34 mm.    Sieve size 3 means pods that fall through a sieve grader which culls    out pod diameters of 7.34 mm through 8.34 mm. Sieve size 4 means    pods that fall through a sieve grader which culls out pod diameters    of 8.34 mm through 9.53 mm. Sieve size 5 means pods that fall    through a sieve grader which culls out pod diameters of 9.53 mm    through 10.72 mm. Sieve size 6 means pods that fall through a sieve    grader that will cull out pod diameters of 10.72 mm or larger.-   “Garden bean Yield” (Tons/Acre)—The yield in tons/acre is the actual    yield of the garden beans at harvest.-   “Plant Height”—Plant height is taken from the top of soil to top    most leaf of the plant and is measured in centimeters.-   “Field holding ability”—A bean plant that has field holding ability    means a plant having pods which remain smooth and retain their color    even after the seed is almost fully developed.-   “Machine harvestable plant”—A machine harvestable bush means a bean    plant that stands with pods off the ground. The pods can be removed    by a machine from the plant without leaves and other plant parts    being harvested.-   “Plant adaptability”—A plant having a good plant adaptability means    a plant that will perform well in different growing conditions and    seasons.-   “Nodes to 1^(st) flower”—This is obtained by counting the node above    the point of cotyledon attachment to the node from which the first    peduncle arises.-   “Peduncle”—A peduncle is the stalk that bears flower(s) and    subsequent pod(s) arising from a node.-   “Node”—A node is the thickened enlargement on a plant. It is where    the stipules, leaf and peduncle arise.-   “RHS”—RHS refers to the Royal Horticultural Society of England which    publishes an official botanical color chart quantitatively    identifying colors according to a defined numbering system, The    chart may be purchased from Royal Hort Society Enterprise Ltd RHS    Garden; Wisley, Woking; Surrey GU236QB, UK.-   “Munsell Color Chart—Munsell Color chart is an alternate botanical    color chart quantitatively identifying color according to a defined    numbering system. It may be purchased through APEDX, 432 Steelhead    Way, Boise, Id. 83704 USA.

DETAILED DESCRIPTION OF THE INVENTION

According to the invention, there is provided a novel bean cultivardesignated SB4355. The garden bean cultivar SB4355 originated from ahand-pollinated cross between Syngenta Seeds, Inc. breeding linesRB3038-P-1-2-2 and RB1320-P-M-16-1-G-1. FIG. 1 provides a detailedpedigree trace for garden been cultivate SB4355.

The mass selection method of breeding was followed during F2 and F3generations to accumulate the traits of upright bush habit, medium greenstraight pods (suitable for processing) and yield. The pedigree methodof selection was utilized in the F4 and F5 generations. The F6generation was bulk harvested to supply a seed source for furtherincreases. SB4355 has been uniform and genetically stable for fivegenerations of increases from the original F6, and is free of off-typesand variants.

Garden bean cultivar SB4355 is a bush snap bean suitable for processingmarkets and can be compared to the variety ‘Hayden’ from Syngenta Seeds,Inc. Garden bean cultivar SB4355 and Hayden are significantly differentin plant height and spread. These measurements were collected from twodifferent locations in Idaho during the 2008 season. ‘Hayden’ was anaverage of 5.25 cm and 3.75 cm taller than SB4355 within the twolocations grown in 2008. Also, the plant spread of variety ‘Hayden’ wasan average of 9.5 cm greater and 7.25 cm greater than the respectiveplant height averages of SB4355 from the 2008 locations.

The values for plant height (cm) and plant spread (cm) are based onmeasurements taken from two different locations grown in Nampa, Id.,during the 2008 growing season. Twenty plants of each cultivar weremeasured for plant height and spread. Each plot was comprised of 100plants.

SB4355 reaches maturity to edible pods in approximately 54 days. Theleaves of SB4355 are medium in size, medium green, and indeterminatesurface. SB4355 anthocyanin pigments are absent from flowers, stems,pods, seeds, leaves, peduncle, petioles and nodes. The color of thewings and keel flower parts of SB4355 are white. The exterior color offresh pods of SB4355 are dark green and the processed pods are dark(Bush Blue Lake 290). The color of the dry pod of SB4355 is buckskin.The pods of SB4355 are low fiber and suture string is absent. Pods fromSB4355 plants contain about six seeds per pod with slow seeddevelopment. Plants and pods of SB4355 are suitable for machine harvest.

Sieve distribution from the two locations grown in 2008 (Nampa, Id.) areas follows:

-   1 Sieve—4.76 to 5.76 mm: 0%-   2 Sieve—5.76 to 7.34 mm: 0%-   3 Sieve—7.34 to 8.34 mm: 18.4%-   4 Sieve—8.34 to 9.53 mm: 33%-   5 Sieve—9.53 to 10.72 mm: 39%-   6 Sieve—>10.72 mm: 9.5%    (Average of two locations grown in Idaho in 2008).

The seed coat luster of SB4355 is semi-shiny, and the seed coat ismonochrome. SB4355 plants primary seed color is white with a white seedcoat pattern. The seeds show a white Hilar ring. One hundred seeds fromSB4355 plants weigh about 18 grams. Plants of SB4355 have a wideadaptation for high yield over a range of environments.

This invention also is directed to methods for producing a garden beanplant by crossing a first parent garden bean plant with a second parentgarden bean plant wherein either the first or second parent garden beanplant is a garden bean plant of the SB4355 line. Still further, thisinvention also is directed to methods for producing a cultivarSB4355-derived garden bean plant by crossing cultivar SB4355 with asecond garden bean plant and growing the progeny seed, and repeating thecrossing and growing steps with the cultivar SB4355-derived plant from 0to 7 times. Thus, any such methods using the cultivar SB4355 are part ofthis invention: selfing, backcrosses, hybrid production, crosses topopulations, and the like. All plants produced using cultivar SB4355 asa parent are within the scope of this invention, including plantsderived from cultivar SB4355. Advantageously, the cultivar is used incrosses with other, different, cultivars to produce first generation(F1) garden bean seeds and plants with superior characteristics.

As used herein, the term plant includes plant cells, plant protoplasts,plant cell tissue cultures from which garden bean plants can beregenerated, plant calli, plant clumps and plant cells that are intactin plants or parts of plants, such as embryos, pollen, ovules, flowers,seeds, pods, stems, roots, anthers, and the like.

With the advent of molecular biological techniques that have allowed theisolation and characterization of genes that encode specific proteinproducts, scientists in the field of plant biology developed a stronginterest in engineering the genome of plants to contain and expressforeign genes, or additional, or modified versions of native, orendogenous, genes (perhaps driven by different promoters) in order toalter the traits of a plant in a specific manner. Such foreignadditional and/or modified genes are referred to herein collectively as“transgenes”. Over the last fifteen to twenty years several methods forproducing transgenic plants have been developed, and the presentinvention, in particular embodiments, also relates to transformedversions of the claimed line.

Plant transformation involves the construction of an expression vectorwhich will function in plant cells. Such a vector comprises DNAcomprising a gene under control of or operatively linked to a regulatoryelement (for example, a promoter). The expression vector may contain oneor more such operably linked gene/regulatory element combinations. Thevector(s) may be in the form of a plasmid, and can be used alone or incombination with other plasmids, to provide transformed garden beanplants, using transformation methods as described below to incorporatetransgenes into the genetic material of the garden bean plant(s).

Expression Vectors for Bean Transformation

Marker Genes—Expression vectors include at least one genetic marker,operably linked to a regulatory element (a promoter, for example) thatallows transformed cells containing the marker to be either recovered bynegative selection, i.e., inhibiting growth of cells that do not containthe selectable marker gene, or by positive selection, i.e., screeningfor the product encoded by the genetic marker. Many commonly usedselectable marker genes for plant transformation are well known in thetransformation arts, and include, for example, genes that code forenzymes that metabolically detoxify a selective chemical agent which maybe an antibiotic or a herbicide, or genes that encode an altered targetwhich is insensitive to the inhibitor. A few positive selection methodsare also known in the art.

One commonly used selectable marker gene for plant transformation is theneomycin phosphotransferase II (nptII) gene, isolated from transposonTn5, which when placed under the control of plant regulatory signalsconfers resistance to kanamycin. Fraley et al., Proc. Natl. Acad. Sci.U.S.A., 80:4803 (1983), Aragao F. J. L., et al., Molecular Breeding 4:6491-499 (1998). Another commonly used selectable marker gene is thehygromycin phosphotransferase gene which confers resistance to theantibiotic hygromycin. Vanden Elzen et al., Plant Mol. Biol., 5:299(1985). Additional selectable marker genes of bacterial origin thatconfer resistance to antibiotics include gentamycin acetyl transferase,streptomycin phosphotransferase, aminoglycoside-3′-adenyl transferase,the bleomycin resistance determinant. Hayford et al., Plant Physiol.86:1216 (1988), Jones et al., Mol. Gen. Genet., 210:86 (1987), Svab etal., Plant Mol. Biol. 14:197 (1990), Hille et al., Plant Mol. Biol.7:171 (1986).

Other selectable marker genes confer resistance to herbicides such asglyphosate, glufosinate or broxynil. Comai et al., Nature 317:741-744(1985), Gordon-Kamm et al., Plant Cell 2:603-618 (1990) and Stalker etal., Science 242:419-423 (1988), Saker M. M., et al, Biologia Plantarum40:4 507-514 (1998), Russel, D. R., et al, Plant Cell Report 12:3165-169 (1993).

Other selectable marker genes for plant transformation are not ofbacterial origin. These genes include, for example, mouse dihydrofolatereductase, plant 5-enolpyruvylshikimate-3-phosphate synthase and plantacetolactate synthase. Eichholtz et al., Somatic Cell Mol. Genet. 13:67(1987), Shah et al., Science 233:478 (1986), Charest et al., Plant CellRep. 8:643 (1990).

Another class of marker genes for plant transformation require screeningof presumptively transformed plant cells rather than direct geneticselection of transformed cells for resistance to a toxic substance suchas an antibiotic. These genes are particularly useful to quantify orvisualize the spatial pattern of expression of a gene in specifictissues and are frequently referred to as reporter genes because theycan be fused to a gene or gene regulatory sequence for the investigationof gene expression. Commonly used genes for screening presumptivelytransformed cells include beta-glucuronidase (GUS), alpha-galactosidase,luciferase and chloramphenicol, acetyltransferase. Jefferson, R. A.,Plant Mol. Biol. Rep. 5:387 (1987), Teeri et al., EMBO J. 8:343 (1989),Koncz et al., Proc. Natl. Acad. Sci U.S.A. 84:131 (1987), DeBlock etal., EMBO J. 3:1681 (1984), Grossi M. F., et al., Plant Science 103:2189-198 (1994), Lewis M. E., Journal of the American Society forHorticultural Science 119:2 361-366 (1994), Zhang et al., Journal of theAmerican Society for Horticultural Science 122:3 300-305 (1997).

Recently, in vivo methods for visualizing GUS activity that do notrequire destruction of plant tissue have been made available. MolecularProbes publication 2908, Imagene Green_, p. 1-4 (1993) and Naleway etal., J. Cell Biol. 115:151a (1991). However, these in vivo methods forvisualizing GUS activity have not proven useful for recovery oftransformed cells because of low sensitivity, high fluorescentbackgrounds and limitations associated with the use of luciferase genesas selectable markers.

More recently, a gene encoding Green Fluorescent Protein (GFP) has beenutilized as a marker for gene expression in prokaryotic and eukaryoticcells. Chalfie et al., Science 263:802 (1994). GFP and mutants of GFPmay be used as screenable markers.

Promoters

Genes included in expression vectors must be driven by nucleotidesequence comprising a regulatory element, for example, a promoter.Several types of promoters are now well known in the transformationarts, as are other regulatory elements that can be used alone or incombination with promoters.

As used herein, “promoter” includes reference to a region of DNAupstream from the start of transcription and involved in recognition andbinding of RNA polymerase and other proteins to initiate transcription.A “plant promoter” is a promoter capable of initiating transcription inplant cells. Examples of promoters under developmental control includepromoters that preferentially initiate transcription in certain tissues,such as leaves, roots, seeds, fibers, xylem vessels, tracheids, orsclerenchyma. Such promoters are referred to as “tissue-preferred”.Promoters which initiate transcription only in certain tissue arereferred to as “tissue-specific”. A “cell type” specific promoterprimarily drives expression in certain cell types in one or more organs,for example, vascular cells in roots or leaves. An “inducible” promoteris a promoter which is under environmental control. Examples ofenvironmental conditions that may effect transcription by induciblepromoters include anaerobic conditions or the presence of light.Tissue-specific, tissue-preferred, cell type specific, and induciblepromoters constitute the class of “non-constitutive” promoters. A“constitutive” promoter is a promoter which is active under mostenvironmental conditions.

A. Inducible Promoters

An inducible promoter is operably linked to a gene for expression ingarden bean. Optionally, the inducible promoter is operably linked to anucleotide sequence encoding a signal sequence which is operably linkedto a gene for expression in garden bean. With an inducible promoter therate of transcription increases in response to an inducing agent.

Any inducible promoter can be used in the instant invention. See Ward etal., Plant Mol. Biol. 22:361-366 (1993). Exemplary inducible promotersinclude, but are not limited to, that from the ACEI system whichresponds to copper (Meft et al., PNAS 90:4567-4571 (1993)); In2 genefrom maize which responds to benzenesulfonamide herbicide safeners(Hershey et al., Mol. Gen Genetics 227:229-237 (1991) and Gatz et al.,Mol. Gen. Genetics 243:32-38 (1994)) or Tet repressor from Tn10 (Gatz etal., Mol. Gen. Genetics 227:229-237 (1991). A particularly preferredinducible promoter is a promoter that responds to an inducing agent towhich plants do not normally respond. An exemplary inducible promoter isthe inducible promoter from a steroid hormone gene, the transcriptionalactivity of which is induced by a glucocorticosteroid hormone. Schena etal., Proc. Natl. Acad. Sci. U.S.A. 88:0421 (1991).

B. Constitutive Promoters

A constitutive promoter is operably linked to a gene for expression ingarden bean or the constitutive promoter is operably linked to anucleotide sequence encoding a signal sequence which is operably linkedto a gene for expression in garden bean.

Many different constitutive promoters can be utilized in the instantinvention. Exemplary constitutive promoters include, but are not limitedto, the promoters from plant viruses such as the 35S promoter from CaMV(Odell et al., Nature 313:810-812 (1985), Aragao et al., Genetics andMolecular Biology 22:3, 445-449 (1999) and the promoters from such genesas rice actin (McElroy et al., Plant Cell 2:163-171 (1990)); ubiquitin(Christensen et al., Plant Mol. Biol. 12:619-632 (1989) and Christensenet al., Plant Mol. Biol. 18:675-689 (1992)); pEMU (Last et al., Theor.Appl. Genet. 81:581-588 (1991)); MAS (Velten et al., EMBO J. 3:2723-2730(1984)) and maize H3 histone (Lepetit et al., Mol. Gen. Genetics231:276-285 (1992) and Atanassova et al., Plant Journal 2 (3): 291-300(1992)).

The ALS promoter, Xba1/Ncol fragment 5′ to the Brassica napus ALS3structural gene (or a nucleotide sequence similarity to said Xba1/Ncolfragment), represents a particularly useful constitutive promoter. SeePCT application WO96/30530.

C. Tissue-Specific or Tissue-Preferred Promoters

A tissue-specific promoter is operably linked to a gene for expressionin garden bean. Optionally, the tissue-specific promoter is operablylinked to a nucleotide sequence encoding a signal sequence which isoperably linked to a gene for expression in garden bean. Plantstransformed with a gene of interest operably linked to a tissue-specificpromoter produce the protein product of the transgene exclusively, orpreferentially, in a specific tissue.

Any tissue-specific or tissue-preferred promoter can be utilized in theinstant invention. Exemplary tissue-specific or tissue-preferredpromoters include, but are not limited to, a root-preferred promoter,such as that from the phaseolin gene (Murai et al., Science 23:476-482(1983) and Sengupta-Gopalan et al., Proc. Natl. Acad. Sci. U.S.A.82:3320-3324 (1985)); a leaf-specific and light-induced promoter such asthat from cab or rubisco (Simpson et al., EMBO J. 4(11):2723-2729 (1985)and Timko et al., Nature 318:579-582 (1985)); an anther-specificpromoter such as that from LAT52 (Twell et al., Mol. Gen. Genetics217:240-245 (1989)); a pollen-specific promoter such as that from Zm13(Guerrero et al., Mol. Gen. Genetics 244:161-168 (1993)) or amicrospore-preferred promoter such as that from apg (Twell et al., Sex.Plant Reprod. 6:217-224 (1993).

Signal Sequences for Targeting Proteins to Subcellular Compartments

Transport of protein produced by transgenes to a subcellular compartmentsuch as the chloroplast, vacuole, peroxisome, glyoxysome, cell wall ormitochondroin or for secretion into the apoplast, is accomplished bymeans of operably linking the nucleotide sequence encoding a signalsequence to the 5′ and/or 3′ region of a gene encoding the protein ofinterest. Targeting sequences at the 5′ and/or 3′ end of the structuralgene may determine, during protein synthesis and processing, where theencoded protein is ultimately compartmentalized.

The presence of a signal sequence directs a polypeptide to either anintracellular organelle or subcellular compartment or for secretion tothe apoplast. Many signal sequences are known in the art. See, forexample Becker et al., Plant Mol. Biol. 20:49 (1992), Close, P. S.,Master's Thesis, Iowa State University (1993), Knox, C., et al.,“Structure and Organization of Two Divergent Alpha-Amylase Genes fromBarley”, Plant Mol. Biol. 9:3-17 (1987), Lerner et al., Plant Physiol.91:124-129 (1989), Fontes et al., Plant Cell 3:483-496 (1991), Matsuokaet al., Proc. Natl. Acad. Sci. 88:834 (1991), Gould et al., J. Cell.Biol. 108:1657 (1989), Creissen et al., Plant J. 2:129 (1991), Kalderon,et al., A short amino acid sequence able to specify nuclear location,Cell 39:499-509 (1984), Steifel, et al., Expression of a maize cell wallhydroxyproline-rich glycoprotein gene in early leaf and root vasculardifferentiation, Plant Cell 2:785-793 (1990).

Foreign Protein Genes and Agronomic Genes

With transgenic plants according to the present invention, a foreignprotein can be produced in commercial quantities. Thus, techniques forthe selection and propagation of transformed plants, which are wellunderstood in the art, yield a plurality of transgenic plants which areharvested in a conventional manner, and a foreign protein then can beextracted from a tissue of interest or from total biomass. Proteinextraction from plant biomass can be accomplished by known methods whichare discussed, for example, by Heney and Orr, Anal. Biochem. 114:92-6(1981).

According to a preferred embodiment, the transgenic plant provided forcommercial production of foreign protein is garden bean. In anotherpreferred embodiment, the biomass of interest is seed. For therelatively small number of transgenic plants that show higher levels ofexpression, a genetic map can be generated, primarily via conventionalRFLP, PCR and SSR analysis, which identifies the approximate chromosomallocation of the integrated DNA molecule. For exemplary methodologies inthis regard, see Glick and Thompson, Methods in Plant Molecular Biologyand Biotechnology CRC Press, Boca Raton 269:284 (1993). Map informationconcerning chromosomal location is useful for proprietary protection ofa subject transgenic plant. If unauthorized propagation is undertakenand crosses made with other germplasm, the map of the integration regioncan be compared to similar maps for suspect plants, to determine if thelatter have a common parentage with the subject plant. Map comparisonswould involve hybridizations, RFLP, PCR, SSR and sequencing, all ofwhich are conventional techniques.

Likewise, by means of the present invention, agronomic genes can beexpressed in transformed plants. More particularly, plants can begenetically engineered to express various phenotypes of agronomicinterest. Exemplary genes implicated in this regard include, but are notlimited to, those categorized below:

1. Genes that Confer Resistance to Pests or Disease and that EncodeEnzymes, Peptides, etc.

-   A. Plant disease resistance genes. Plant defenses are often    activated by specific interaction between the product of a disease    resistance gene (R) in the plant and the product of a corresponding    avirulence (Avr) gene in the pathogen. A plant line can be    transformed with cloned resistance gene to engineer plants that are    resistant to specific pathogen strains. See, for example Jones et    al., Science 266:789 (1994) (cloning of the tomato Cf-9 gene for    resistance to Cladosporium fulvum); Martin et al., Science    262:1432 (1993) (tomato Pto gene for resistance to Pseudomonas    syringae pv. Tomato encodes a protein kinase); Mindrinos et al.,    Cell 78:1089 (1994) (Arabidopsis RSP2 gene for resistance to    Pseudomonas syingae).-   B. A Bacillus thuringiensis protein, a derivative thereof or a    synthetic polypeptide modeled thereon. See, for example, Geiser et    al., Gene 48:109 (1986), who disclose the cloning and nucleotide    sequence of a Bt delta-endotoxin gene. Moreover, DNA molecules    encoding delta-endotoxin genes can be purchased from American Type    Culture Collection, Manassas, Va., for example, under ATCC Accession    Nos. 40098, 67136, 31995 and 31998.-   C. A lectin. See, for example, the disclose by Van Damme et al.,    Plant Molec. Biol. 24:25 (1994), who disclose the nucleotide    sequences of several Clivia miniata mannose-binding lectin genes.-   D. A vitamin-binding protein such as avidin. See PCT application    US93/06487, the contents of which are hereby incorporated by    reference. The application teaches the use of avidin and avidin    homologues as larvicides against insect pests.-   E. An enzyme inhibitor, for example, a protease or proteinase    inhibitor or an amylase inhibitor. See, for example, Abe et al., J.    Biol. Chem. 262:16793 (1987) (nucleotide sequence of rice cysteine    proteinase inhibitor), Huub et al., Plant Molec. Biol. 21:985 (1993)    (nucleotide sequence of cDNA encoding tobacco proteinase inhibitor    I), Sumitani et al., Biosci. Biotech. Biochem. 57:1243 (1993)    (nucleotide sequence of Streptomyces nitrosporeus alpha-amylase    inhibitor).-   F. An insect-specific hormone or pheromone such as an ecdysteroid    and juvenile hormone, a variant thereof, a mimetic based thereon, or    an antagonist or agonist thereof. See, for example, the disclosure    by Hammock et al., Nature 344:458 (1990), of baculovirus expression    of cloned juvenile hormone esterase, an inactivator of juvenile    hormone.-   G. An insect-specific peptide or neuropeptide which, upon    expression, disrupts the physiology of the affected pest. For    example, see the disclosures of Regan, J. Biol. Chem. 269:9 (1994)    (expression cloning yields DNA coding for insect diuretic hormone    receptor), and Pratt et al., Biochem. Biophys. Res. Comm.    163:1243 (1989) (an allostatin is identified in Diploptera    punctata). See also U.S. Pat. No. 5,266,317 to Tomalski et al., who    disclose genes encoding insect-specific, paralytic neurotoxins.-   H. An insect-specific venom produced in nature by a snake, a wasp,    etc. For example, see Pang et al., Gene 116:165 (1992), for    disclosure of heterologous expression in plants of a gene coding for    a scorpion insectotoxic peptide.-   I. An enzyme responsible for a hyper accumulation of a monterpene, a    sesquiterpene, a steroid, hydroxamic acid, a phenylpropanoid    derivative or another non-protein molecule with insecticidal    activity.-   J. An enzyme involved in the modification, including the    post-translational modification, of a biologically active molecule;    for example, a glycolytic enzyme, a proteolytic enzyme, a lipolytic    enzyme, a nuclease, a cyclase, a transaminase, an esterase, a    hydrolase, a phosphatase, a kinase, a phosphorylase, a polymerase,    an elastase, a chitinase and a glucanase, whether natural or    synthetic. See PCT application WO 93/02197 in the name of Scott et    al., which discloses the nucleotide sequence of a callase gene. DNA    molecules which contain chitinase-encoding sequences can be    obtained, for example, from the ATCC under Accession Nos. 39637    and 67152. See also Kramer et al., Insect Biochem. Molec. Biol.    23:691 (1993), who teach the nucleotide sequence of a cDNA encoding    tobacco hookworm chitinase, and Kawalleck et al., Plant Molec. Biol.    21:673 (1993), who provide the nucleotide sequence of the parsley    ubi4-2 polyubiquitin gene.-   K. A molecule that stimulates signal transduction. For example, see    the disclosure by Botella et al., Plant Molec. Biol. 24:757 (1994),    of nucleotide sequences for mung bean calmodulin cDNA clones, and    Griess et al., Plant Physiol. 104:1467 (1994), who provide the    nucleotide sequence of a maize calmodulin cDNA clone.-   L. A hydrophobic moment peptide. See PCT application WO95/16776    (disclosure of peptide derivatives of Tachyplesin which inhibit    fungal plant pathogens) and PCT application WO95/18855 (teaches    synthetic antimicrobial peptides that confer disease resistance),    the respective contents of which are hereby incorporated by    reference.-   M. A membrane permease, a channel former or a channel blocker. For    example, see the disclosure of Jaynes et al., Plant Sci 89:43    (1993), of heterologous expression of a cecropin-beta, lytic peptide    analog to render transgenic tobacco plants resistant to Pseudomonas    solanacearum.-   N. A viral-invasive protein or a complex toxin derived therefrom.    For example, the accumulation of viral coat proteins in transformed    plant cells imparts resistance to viral infection and/or disease    development effected by the virus from which the coat protein gene    is derived, as well as by related viruses. See Beachy et al., Ann.    rev. Phytopathol. 28:451 (1990). Coat protein-mediated resistance    has been conferred upon transformed plants against alfalfa mosaic    virus, cucumber mosaic virus, tobacco streak virus, potato virus X,    potato virus Y, tobacco etch virus, tobacco rattle virus and tobacco    mosaic virus. Id.-   O. An insect-specific antibody or an immunotoxin derived therefrom.    Thus, an antibody targeted to a critical metabolic function in the    insect gut would inactivate an affected enzyme, killing the insect.    Cf. Taylor et al., Abstract #497, Seventh Int'l Symposium on    Molecular Plant-Microbe Interactions (Edinburgh, Scotland) (1994)    (enzymatic inactivation in transgenic tobacco via production of    single-chain antibody fragments).-   P. A virus-specific antibody. See, for example, Tavladoraki et al.,    Nature 366:469 (1993), who show that transgenic plants expressing    recombinant antibody genes are protected from virus attack.-   Q. A developmental-arrestive protein produced in nature by a    pathogen or a parasite. Thus, fungal endo    alpha-1,4-D-polygalacturonases facilitate fungal colonization and    plant nutrient release by solubilizing plant cell wall    homo-alpha-1,4-D-galacturonase. See Lamb et al., Bio/Technology    10:1436 (1992). The cloning and characterization of a gene which    encodes a bean endopolygalacturonase-inhibiting protein is described    by Toubart et al., Plant J. 2:367 (1992).-   R. A development-arrestive protein produced in nature by a plant.    For example, Logemann et al., Bioi/Technology 10:305 (1992), have    shown that transgenic plants expressing the barley    ribosome-inactivating gene have an increased resistance to fungal    disease.    2. Genes that Confer Resistance to a Herbicide, for Example-   A. A herbicide that inhibits the growing point or meristem, such as    an imidazalinone or a sulfonylurea. Exemplary genes in this category    code for mutant ALS and AHAS enzyme as described, for example, by    Lee et al., EMBO J. 7:1241 (1988), and Miki et al., Theor. Appl.    Genet. 80:449 (1990), respectively.-   B. Glyphosate (resistance impaired by mutant    5-enolpyruvl-3-phosphikimate synthase (EPSP) and aroA genes,    respectively) and other phosphono compounds such as glufosinate    (phosphinothricin acetyl transferase, PAT and Streptomyces    hygroscopicus phosphinothricin-acetyl transferase, bar, genes), and    pyridinoxy or phenoxy propionic acids and cycloshexones (ACCase    inhibitor-encoding genes). See, for example, U.S. Pat. No. 4,940,835    to Shah, et al., which discloses the nucleotide sequence of a form    of EPSP which can confer glyphosate resistance. A DNA molecule    encoding a mutant aroA gene can be obtained under ATCC accession    number 39256, and the nucleotide sequence of the mutant gene is    disclosed in U.S. Pat. No. 4,769,061 to Comai. European patent    application No. 0 333 033 to Kumada et al., and U.S. Pat. No.    4,975,374 to Goodman et al., disclose nucleotide sequences of    glutamine synthetase genes which confer resistance to herbicides    such as L-phosphinothricin. See also Russel, D. R., et al, Plant    Cell Report 12:3 165-169 (1993). The nucleotide sequence of a    phosphinothricin-acetyl-transferase gene is provided in European    application No. 0 242 246 to Leemans et al., DeGreef et al.,    Bio/Technology 7:61 (1989), describe the production of transgenic    plants that express chimeric bar genes coding for phosphinothricin    acetyl transferase activity. Exemplary of genes conferring    resistance to phenoxy propionic acids and cycloshexones, such as    sethoxydim and haloxyfop are the Acc1-S1, Acc1-S2 and Acc1-S3 genes    described by Marshall et al., Theor. Appl. Genet. 83:435 (1992).-   C. A herbicide that inhibits photosynthesis, such as a triazine    (psbA and gs+ genes) and a benzonitrile (nitrilase gene). Przibilla    et al., Plant Cell 3:169 (1991), describe the transformation of    Chlamydomonas with plasmids encoding mutant psbA genes. Nucleotide    sequences for nitrilase genes are disclosed in U.S. Pat. No.    4,810,648 to Stalker, and DNA molecules containing these genes are    available under ATCC Accession Nos. 53435, 67441, and 67442. Cloning    and expression of DNA coding for a glutathione S-transferase is    described by Hayes et al., Biochem. J. 285:173 (1992).    3. Genes that Confer or Contribute to a Value-Added Trait, such as-   A. Delayed and attenuated symptoms to Bean Golden Mosaic Geminivirus    (BGMV), for example by transforming a plant with antisense genes    from the Brazilian BGMV. See Arago et al., Molecular Breeding. 1998,    4: 6, 491-499.-   B. Increased the pea content in Methionine by introducing a    transgene coding for a Methionine rich storage albumin (2S-albumin)    from the Brazil nut as decribed in Arago et al., Genetics and    Molecular Biology. 1999, 22: 3, 445-449.-   Numerous methods for plant transformation have been developed,    including biological and physical, plant transformation protocols.    See, for example, Miki et al., “Procedures for Introducing Foreign    DNA into Plants” in Methods in Plant Molecular Biology and    Biotechnology, Glick B. R. and Thompson, J. E. Eds. (CRC Press,    Inc., Boca Raton, 1993) pages 67-88. In addition, expression vectors    and in vitro culture methods for plant cell or tissue transformation    and regeneration of plants are available. See, for example, Gruber    et al., “Vectors for Plant Transformation” in Methods in Plant    Molecular Biology and Biotechnology, Glick B. R. and Thompson, J. E.    Eds. (CRC Press, Inc., Boca Raton, 1993) pages 89-119.

A. Agrobacterium-Mediated Transformation

One method for introducing an expression vector into plants is based onthe natural transformation system of Agrobacterium. See, for example,Horsch et al., Science 227:1229 (1985). McClean, P., et al. Plant CellTissue Org. Cult. 24(2, February), 131-138 (1991), Lewis et al., Journalof the American Society for Horticultural Science, 119:2, 361-366(1994), Zhang, Z., et al. J. Amer. Soc. Hort. Sci. 122(3): 300-305(1997). A. tumefaciens and A. rhizogenes are plant pathogenic soilbacteria which genetically transform plant cells. The Ti and Ri plasmidsof A. tumefaciens and A. rhizogenes, respectively, carry genesresponsible for genetic transformation of the plant. See, for example,Kado, C. I., Crit. Rev. Plant Sci. 10:1 (1991). Descriptions ofAgrobacterium vector systems and methods for Agrobacterium-mediated genetransfer are provided by Gruber et al., supra, Miki et al., supra, andMoloney et al., Plant Cell Reports 8:238 (1989). See also, U.S. Pat. No.5,591,616 issued Jan. 7, 1997.

B. Direct Gene Transfer

Despite the fact the host range for Agrobacterium-mediatedtransformation is broad, some major cereal or vegetable crop species andgymnosperms have generally been recalcitrant to this mode of genetransfer, even though some success has recently been achieved in riceand corn. Hiei et al., The Plant Journal 6:271-282 (1994) and U.S. Pat.No. 5,591,616 issued Jan. 7, 1997. Several methods of planttransformation, collectively referred to as direct gene transfer, havebeen developed as an alternative to Agrobacterium-mediatedtransformation.

A generally applicable method of plant transformation ismicroprojectile-mediated transformation wherein DNA is carried on thesurface of microprojectiles measuring 1 to 4 im. The expression vectoris introduced into plant tissues with a biolistic device thataccelerates the microprojectiles to speeds of 300 to 600 m/s which issufficient to penetrate plant cell walls and membranes. Russell, D. R.,et al. PI. Cell. Rep. 12(3, January), 165-169 (1993), Aragao, F. J. L.,et al. Plant Mol. Biol. 20(2, October), 357-359 (1992), Aragao Theor.Appl. Genet. 93:142-150 (1996), Kim, J.; Minamikawa, T. Plant Science117: 131-138 (1996), Sanford et al., Part. Sci. Technol. 5:27 (1987),Sanford, J. C., Trends Biotech. 6:299 (1988), Klein et al.,Bio/Technology 6:559-563 (1988), Sanford, J. C., Physiol Plant 7:206(1990), Klein et al., Biotechnology 10:268 (1992)

Another method for physical delivery of DNA to plants is sonication oftarget cells. Zhang et al., Bio/Technology 9:996 (1991). Alternatively,liposome or spheroplast fusion have been used to introduce expressionvectors into plants. Deshayes et al., EMBO J., 4:2731 (1985), Christouet al., Proc Natl. Acad. Sci. U.S.A. 84:3962 (1987). Direct uptake ofDNA into protoplasts using CaCl₂ precipitation, polyvinyl alcohol orpoly-L-omithine have also been reported. Hain et al., Mol. Gen. Genet.199:161 (1985) and Draperetal., Plant Cell Physiol. 23:451 (1982).Electroporation of protoplasts and whole cells and tissues have alsobeen described. Saker, M.; Kuhne, T. Biologia Plantarum 40(4): 507-514(1997/98), Donn et al., In Abstracts of VIIth International Congress onPlant Cell and Tissue Culture IAPTC, A2-38, p 53 (1990); D'Halluin etal., Plant Cell 4:1495-1505 (1992) and Spencer et al., Plant Mol. Biol.24:51-61 (1994).

Following transformation of garden bean target tissues, expression ofthe above-described selectable marker genes allows for preferentialselection of transformed cells, tissues and/or plants, usingregeneration and selection methods now well known in the art.

The foregoing methods for transformation would typically be used forproducing a transgenic line. The transgenic line could then be crossed,with another (non-transformed or transformed) line, in order to producea new transgenic garden bean line. Alternatively, a genetic trait whichhas been engineered into a particular garden bean cultivar using theforegoing transformation techniques could be moved into another lineusing traditional backcrossing techniques that are well known in theplant breeding arts. For example, a backcrossing approach could be usedto move an engineered trait from a public, non-elite inbred line into anelite inbred line, or from an inbred line containing a foreign gene inits genome into an inbred line or lines which do not contain that gene.As used herein, “crossing” can refer to a simple X by Y cross, or theprocess of backcrossing, depending on the context.

When the term garden bean plant, cultivar or garden bean line is used inthe context of the present invention, this also includes any single geneconversions of that cultivar or line. The term single gene convertedplant as used herein refers to those garden bean plants which aredeveloped by a plant breeding technique called backcrossing whereinessentially all of the desired morphological and physiologicalcharacteristics of a cultivar are recovered in addition to the singlegene transferred into the line via the backcrossing technique.Backcrossing methods can be used with the present invention to improveor introduce a characteristic into the line. The term backcrossing asused herein refers to the repeated crossing of a hybrid progeny back toone of the parental garden bean plants for that line. The parentalgarden bean plant which contributes the gene for the desiredcharacteristic is termed the nonrecurrent or donor parent. Thisterminology refers to the fact that the nonrecurrent parent is used onetime in the backcross protocol and therefore does not recur. Theparental garden bean plant to which the gene or genes from thenonrecurrent parent are transferred is known as the recurrent parent asit is used for several rounds in the backcrossing protocol (Poehlman &Sleper, 1994; Fehr, 1987). In a typical backcross protocol, the originalcultivar of interest (recurrent parent) is crossed to a second line(nonrecurrent parent) that carries the single gene of interest to betransferred. The resulting progeny from this cross are then crossedagain to the recurrent parent and the process is repeated until a gardenbean plant is obtained wherein essentially all of the desiredmorphological and physiological characteristics of the recurrent parentare recovered in the converted plant, in addition to the singletransferred gene from the nonrecurrent parent.

The selection of a suitable recurrent parent is an important step for asuccessful backcrossing procedure. The goal of a backcross protocol isto alter or substitute a single trait or characteristic in the originalline. To accomplish this, a single gene of the recurrent cultivar ismodified or substituted with the desired gene from the nonrecurrentparent, while retaining essentially all of the rest of the desiredgenetic, and therefore the desired physiological and morphological,constitution of the original line. The choice of the particularnonrecurrent parent will depend on the purpose of the backcross, one ofthe major purposes is to add some commercially desirable, agronomicallyimportant trait to the plant. The exact backcrossing protocol willdepend on the characteristic or trait being altered to determine anappropriate testing protocol. Although backcrossing methods aresimplified when the characteristic being transferred is a dominantallele, a recessive allele may also be transferred. In this instance itmay be necessary to introduce a test of the progeny to determine if thedesired characteristic has been successfully transferred.

Many single gene traits have been identified that are not regularlyselected for in the development of a new line but that can be improvedby backcrossing techniques. Single gene traits may or may not betransgenic, examples of these traits include but are not limited to,herbicide resistance (such as bar or pat genes), resistance forbacterial, fungal, or viral disease such as gene I used for BCMVresistance), insect resistance, enhanced nutritional quality (such as 2salbumine gene), industrial usage, agronomic qualities such as the“persistent green gene”, yield stability and yield enhancement. Thesegenes are generally inherited through the nucleus. Some other singlegene traits are described in U.S. Pat. Nos. 5,777,196; 5,948,957 and5,969,212, the disclosures of which are specifically hereby incorporatedby reference.

EXAMPLES Tables

In the tables 1-5 that follow, the traits and characteristics of gardenbean line SB4355 are given along with data on the commercial garden beanvariety used as a check. Table 6 is the pedigree trace for SB4355.

TABLE 1 Explanation of statistical analysis variables used in thefollowing attachments. Hay_PLTHT1 = Hayden plant height (cm), Location1, Nampa, ID, 2008 Pen_PLTHT1 = SB4355 plant height (cm), Location 1,Nampa, ID, 2008 Hay_PLTHT2 = Hayden plant height (cm), Location 2,Nampa, ID, 2008 Pen_PLTHT2 = SB4355 plant height (cm), Location 2,Nampa, ID, 2008 Hay_PLTSP1 = Hayden plant spread (cm), Location 1,Nampa, ID, 2008 Pen_PLTSP1 = SB4355 plant spread (cm), Location 1,Nampa, ID, 2008 Hay_PLTSP2 = Hayden plant spread (cm), Location 2,Nampa, ID, 2008 Pen_PLTSP2 = SB4355 plant spread (cm), Location 2,Nampa, ID, 2008

TABLE 2 Comparison of plant height (cm) values from Location 1, Nampa,ID, 2008 Descriptive Statistics Variable N Mean SD Minimum MaximumHay_PLTHT1 20 50.750 4.9404 40.000 60.000 Pen_PLTHT1 20 45.500 4.559540.000 50.000 One-Way AOV for: Hay_PLTHT1 Pen_PLTHT1 Source DF SS MS F PBetween 1 275.63 275.625 12.20 0.0012 Within 38 858.75 22.599 Total 391134.37 Grand Mean 48.125 CV  9.88 Homogeneity of Variances F P Levene'sTest 0.19 0.6686 O'Brien's Test 0.18 0.6770 Brown and Forsythe Test 0.620.4353 Welch's Test for Mean Differences Source DF F P Between 1.0 12.200.0012 Within 37.8 Component of variance for between groups 12.6513Effective cell size 20.0 Variable Mean Hay_PLTHT1 50.750 Pen_PLTHT145.500 Observations per Mean 20 Standard Error of a Mean 1.0630 StdError (Diff of 2 Means) 1.5033 LSD All-Pairwise Comparisons TestVariable Mean Homogeneous Groups Hay_PLTHT1 50.750 A Pen_PLTHT1 45.500 BAlpha 0.05 Standard Error for Comparison 1.5033 Critical T Value 2.024Critical Value for Comparison 3.0432 All 2 means are significantlydifferent from one another.

TABLE 3 Comparison of plant height (cm) values from Location 2, Nampa,ID, 2008 Descriptive Statistics Variable N Mean SD Minimum MaximumHay_PLTHT2 20 48.750 4.5523 40.000 60.000 Pen_PLTHT2 20 45.000 4.866640.000 50.000 One-Way AOV for: Hay_PLTHT2 Pen_PLTHT2 Source DF SS MS F PBetween 1 140.625 140.625 6.33 0.0162 Within 38 843.750 22.204 Total 39984.375 Grand Mean 46.875 CV 10.05 Homogeneity of Variances F P Levene'sTest 0.13 0.7224 O'Brien's Test 0.12 0.7296 Brown and Forsythe Test 3.650.0636 Welch's Test for Mean Differences Source DF F P Between 1.0 6.330.0162 Within 37.8 Component of variance for between groups  5.92105Effective cell size 20.0 Variable Mean Hay_PLTHT2 48.750 Pen_PLTHT245.000 Observations per Mean 20 Standard Error of a Mean 1.0537 StdError (Diff of 2 Means) 1.4901 LSD All-Pairwise Comparisons TestVariable Mean Homogeneous Groups Hay_PLTHT2 48.750 A Pen_PLTHT2 45.000 BAlpha 0.05 Standard Error for Comparison 1.4901 Critical T Value 2.024Critical Value for Comparison 3.0165 All 2 means are significantlydifferent from one another.

TABLE 4 Comparison of plant spread (cm) values from Location 1, Nampa,ID, 2008 Descriptive Statistics Variable N Mean SD Minimum MaximumHay_PLTSP1 20 51.250 4.5523 45.000 60.000 Pen_PLTSP1 20 41.750 5.683530.000 50.000 One-Way AOV for: Hay_PLTSP1 Pen_PLTSP1 Source DF SS MS F PBetween 1 902.50 902.500 34.04 0.0000 Within 38 1007.50 26.513 Total 391910.00 Grand Mean 46.500 CV 11.07 Homogeneity of Variances F P Levene'sTest 0.87 0.3579 O'Brien's Test 0.82 0.3709 Brown and Forsythe Test 0.570.4552 Welch's Test for Mean Differences Source DF F P Between 1.0 34.040.0000 Within 36.3 Component of variance for between groups 43.7993Effective cell size 20.0 Variable Mean Hay_PLTSP1 51.250 Pen_PLTSP141.750 Observations per Mean 20 Standard Error of a Mean 1.1514 StdError (Diff of 2 Means) 1.6283 LSD All-Pairwise Comparisons TestVariable Mean Homogeneous Groups Hay_PLTSP1 51.250 A Pen_PLTSP1 41.750 BAlpha 0.05 Standard Error for Comparison 1.6283 Critical T Value 2.024Critical Value for Comparison 3.2963 All 2 means are significantlydifferent from one another.

TABLE 5 Comparison of plant spread (cm) values from Location 2, Nampa,ID, 2008 Descriptive Statistics Variable N Mean SD Minimum MaximumHay_PLTSP2 20 48.250 6.5444 40.000 60.000 Pen_PLTSP2 20 41.000 11.98710.000 50.000 One-Way AOV for: Hay_PLTSP2 Pen_PLTSP2 Source DF SS MS F PBetween 1 525.62 525.625 5.64 0.0227 Within 38 3543.75 93.257 Total 394069.38 Grand Mean 44.625 CV 21.64 Homogeneity of Variances F P Levene'sTest 2.20 0.1462 O'Brien's Test 2.08 0.1571 Brown and Forsythe Test 1.220.2756 Welch's Test for Mean Differences Source DF F P Between 1.0 5.640.0243 Within 29.4 Component of variance for between groups 21.6184Effective cell size 20.0 Variable Mean Hay_PLTSP2 48.250 Pen_PLTSP241.000 Observations per Mean 20 Standard Error of a Mean 2.1594 StdError (Diff of 2 Means) 3.0538 LSD All-Pairwise Comparisons TestVariable Mean Homogeneous Groups Hay_PLTSP2 48.250 A Pen_PLTSP2 41.000 BAlpha 0.05 Standard Error for Comparison 3.0538 Critical T Value 2.024Critical Value for Comparison 6.1821 All 2 means are significantlydifferent from one another.

Deposit

Applicants have made a deposit of at least 2500 seeds of with theAmerican Type Culture Collection (ATCC), Manassas, Va., 20110-2209U.S.A., ATCC Deposit No: ______. This deposit of the garden beancultivar designated SB4355 will be maintained in the ATCC depository,which is a public depository, for a period of 30 years, or 5 years afterthe most recent request, or for the effective life of the patent,whichever is longer, and will be replaced if it becomes nonviable duringthat period. Additionally, Applicants have satisfied all therequirements of 37 C.F.R. §§1.801-1.809, including providing anindication of the viability of the sample. Applicants impose norestrictions on the availability of the deposited material from theATCC; however, Applicants have no authority to waive any restrictionsimposed by law on the transfer of biological material or itstransportation in commerce. Applicants do not waive any infringement ofits rights granted under this patent or under the Plant CultivarProtection Act (7 USC 2321 et seq.).

The foregoing invention has been described in detail by way ofillustration and example for purposes of clarity and understanding.However, it will be obvious that certain changes and modifications suchas single gene modifications and mutations, somaclonal variants, variantindividuals selected from large populations of the plants of the instantinbred and the like may be practiced within the scope of the invention,as limited only by the scope of the appended claims.

All references cited herein are incorporated by reference in the instantapplication in their entireties.

1. Seed of garden bean cultivar designated SB4355, representative seedof said cultivar having been deposited under ATCC Accession No.______.2. The garden bean plant, or a part thereof, produced by growing theseed of claim
 1. 3. Pollen of the plant of claim
 2. 4. An ovule of theplant of claim
 2. 5. A pod or a berry of the plant of claim
 2. 6. Atissue culture of regenerable cells of a plant of garden bean cultivardesignated SB4355, wherein the tissue regenerates plants having all themorphological and physiological characteristics of a plant of gardenbean cultivar designated SB4355, representative seeds having beendeposited ATCC Accession No.______.
 7. The tissue culture of claim 6,selected from the group consisting of protoplast and calli, wherein theregenerable cells are produced from meristematic cells, leaves, pollen,embryo, root, root tips, stems, anther, flowers, seeds or pods.
 8. Agarden bean plant regenerated from the tissue culture of claim 6,wherein the regenerated plant has all the morphological andphysiological characteristics of a plant of garden bean cultivardesignated SB4355, representative seeds having been deposited under ATCCAccession No.______.
 9. A method for producing a hybrid garden bean seedcomprising crossing a first parent garden bean plant with a secondparent garden bean plant and harvesting the resultant hybrid garden beanseed, wherein said first or second parent garden bean plant is a gardenbean plant of claim
 2. 10. The hybrid garden bean seed produced by themethod of claim
 9. 11. A method of producing an herbicide resistantgarden bean plant comprising transforming the garden bean plant of claim2 with a transgene that confers herbicide resistance.
 12. An herbicideresistant garden bean plant produced by the method of claim
 11. 13. Thegarden bean plant of claim 12, wherein the transgene confers resistanceto an herbicide selected from the group consisting of imidazolinone,sulfonylurea glyphosate, glufosinate, L-phosphinothricin, triazine andbenzonitrile.
 14. A method of producing an insect resistant garden beanplant comprising transforming the garden bean plant of claim 2 with atransgene that confers insect resistance.
 15. An insect resistant gardenbean plant produced by the method of claim
 14. 16. The garden bean plantof claim 15, wherein the transgene encodes a Bacillus thuringiensisprotein.
 17. A method of producing a disease resistant garden bean plantcomprising transforming the garden bean plant of claim 2 with atransgene that confers resistance to bacterial, fungal or viral disease.18. A disease resistant garden bean plant produced by the method ofclaim
 17. 19. A method of producing a garden bean pod comprising: a.growing a garden bean plant of claim 2 to produce a garden bean pod, andb. harvesting said garden bean pod.
 20. A method according to claim 19,further comprising processing said garden bean pod to obtain a seed. 21.A method according to claim 20, wherein said seed is a fresh product, acanned product or a frozen product.
 22. A method of producing a seedcomprising obtaining a pod of a plant of claim 2 and processing said podto obtain a seed.
 23. A method according to claim 22, wherein said seedis a fresh product, a canned product or a frozen product.